AFM force measurements of the gp120–sCD4 and gp120 or CD4 antigen–antibody interactions

Soluble CD4 (sCD4), anti-CD4 antibody, and anti-gp120 antibody have long been regarded as entry inhibitors in human immunodeficiency virus (HIV) therapy. However, the interactions between these HIV entry inhibitors and corresponding target molecules are still poorly understood. In this study, atomic force microscopy (AFM) was utilized to investigate the interaction forces among them. We found that the unbinding forces of sCD4–gp120 interaction, CD4 antigen–antibody interaction, and gp120 antigen–antibody interaction were 25.45 ± 20.46, 51.22 ± 34.64, and 89.87 ± 44.63 pN, respectively, which may provide important mechanical information for understanding the effects of viral entry inhibitors on HIV infection. Moreover, we found that the functionalization of an interaction pair on AFM tip or substrate significantly influenced the results, implying that we must perform AFM force measurement and analyze the data with more caution.

► The unbinding force of sCD4–gp120 interaction was 25.45 ± 20.46 pN.
► The unbinding force of CD4 antigen–antibody interaction was 51.22 ± 34.64 pN.
► The unbinding force of gp120 antigen–antibody interaction was 89.87 ± 44.63 pN.
► The interaction forces between various HIV inhibitors and the target molecules are significantly different.
► Functionalizing on AFM tip or substrate of an interaction pair caused different results.